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MTAP - HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES MODULE > LESSON 6: INFILTRATION AND EMBEDDING > Flashcards

LESSON 6: INFILTRATION AND EMBEDDING Flashcards

1
Q

✓ to fill all natural cavities, spaces & interstices of the tissues

A

IMPREGNATION

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2
Q

✓ to give tissue samples a firm consistency

A

IMPREGNATION

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3
Q

✓ impregnated tissue is placed into a precisely arranged position in a mold containing a medium which is then allowed to solidify

A

EMBEDDING

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4
Q

mechanisms of solidification:

A

→crystallization
→evaporation of the solvent
→polymerization

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5
Q

✓ Simplest, most common & best embedding medium

A

PARAFFIN PROCESSING

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6
Q

Methods of Paraffin Impregnation

A
  1. MANUAL PROCESSING
  2. AUTOMATIC PROCESSING
  3. VACUUM EMBEDDING
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7
Q

Substitutes for Paraffin Wax

A
  1. Paraplast
  2. Ester wax
  3. Water Soluble Waxes
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8
Q
  1. Paraplast:
A

Embeddol

Bioloid

Tissue Mat

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9
Q

EMBEDDING METHODS

A

WET METHOD

DRY METHOD

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10
Q

TYPES OF PLASTIC MEDIA

A
  1. EPOXY 2. POLYESTER 3. ACRYLIC
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11
Q
  1. EPOXY:
A

➢ Araldite base (bisphenol) - slowest
➢ Glycerol base (epon)
➢ Cyclohexene Dioxide (spurr) - fastest

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12
Q

Types of Embedding Molds

A
  1. Leuckhart’s embedding mold
  2. Compound E unit
  3. Disposable Molds
  4. Plastic Embedding Rings & Base Molds
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13
Q

Disposable Molds:

A

a. Peel-away (thin plastic Embedding molds)
b. Plastic Ice Trays
c. Paper boats

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14
Q

➢ substance used to infiltrate, support and enclose tissue specimen

A

Infiltration/Embedding media

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15
Q

➢ should be the same for infiltration and embedding

A

Infiltration/Embedding media

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16
Q

Infiltration/Embedding media ➢ most important characteristic:

A

convertible from liquid to solid form

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17
Q

✓ Simplest, most common & best embedding medium

A

PARAFFIN PROCESSING

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18
Q

Clearing Agent Used

A

✓ xylene/benzene vs chloroform/cedarwood oil

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19
Q

❖ 4 changes at 15-minute intervals

A
  1. MANUAL PROCESSING
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20
Q

❖ 2-5 OC higher than the MP of the wax

A
  1. MANUAL PROCESSING
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21
Q

❖ 2 – 3 changes with agitation

A
  1. AUTOMATIC PROCESSING
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22
Q

❖ At least 3 OC higher than the MP of the wax

A
  1. AUTOMATIC PROCESSING
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23
Q

❖ negative atmospheric pressure (400-500 mmHg)

A
  1. VACUUM EMBEDDING
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24
Q

❖ Heat & vacuum

A
  1. VACUUM EMBEDDING
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25
Q

❖ 3 changes

A
  1. VACUUM EMBEDDING
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26
Q

❖ 2 - 4 OC above MP of wax

A
  1. VACUUM EMBEDDING
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27
Q

❖ ADV: Effects of heat are prevented

A
  1. VACUUM EMBEDDING
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28
Q

❖ Applications in the lab: urgent biopsies, dense and fibrous tissues, delicate tissues

A
  1. VACUUM EMBEDDING
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29
Q

➢ Fresh was should be filtered at

A

2 OC above its MP

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30
Q

➢ Wax from trimmings should be filtered with a

A

course filter paper

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31
Q

➢ Water is removed by heating the wax to

A

100-105 OC

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32
Q

✓ Paraffin wax should be

A

pure

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33
Q

Paraffin wax may be utilized

A

twice

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34
Q

✓ Melted paraffin: (?)above the MP of wax
✓ Cooling ➢ Ref at (?)
➢ immerse in cold water
➢ Tissue Tek with cold plate

A

5-10 OC

-5 OC

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35
Q

➢ MP: 56-57 OC

A

Paraplast

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36
Q

➢ More elastic & resilient

A

Paraplast

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37
Q

➢ Doesn’t require cooling

A

Paraplast

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38
Q

➢ MP= 56 – 58 OC

A

❖ Embeddol

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39
Q

➢ Less brittle than paraplast

A

❖ Embeddol

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40
Q

➢ Semisynthetic wax that is used in impregnating and embedding eye samples

A

❖ Bioloid

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41
Q

➢ Product of paraffin that contains rubber

A

❖ Tissue Mat

42
Q

➢ MP: 46 – 48 OC

A

Ester wax

43
Q

➢ Lower melting point but harder than paraffin

A

Ester wax

44
Q

➢ Soluble in 95% EA & other clearing agents

A

Ester wax

45
Q

Ester wax ➢ PROCESSING:

A

cellosolve/xylene → equal parts clearing agent + ester wax for 36 hours → 3-4 changes of pure ester wax → embedding → sectioning using heavy duty microtome

46
Q

➢ Polyethylene glycols

A

Water Soluble Waxes

47
Q

➢ Does not require dehydration & clearing

A

Carbowax

48
Q

Fixation → wash-out → carbowax processing

A

Carbowax

49
Q

✓ Reduces the processing time

A

Carbowax

50
Q

➢ Does not remove neutral fats & lipids

A

Carbowax

51
Q

➢ For histochemical and enzyme studies

A

Carbowax

52
Q

➢ 70 % (?) → 90% (?) → 100% (2 changes for [?]) at 56 OC

A

30 min

45 min

1 hr each

53
Q

➢ Blocking at 50 OC →rapidly cooled at ref temp

A

Carbowax

54
Q

➢ Applications: histochemical and enzyme studies; for the demonstration of neutral fats and lipids

A

Carbowax

55
Q

➢ DISADVANTAGE: hygroscopic reagent

A

Carbowax

56
Q

Carbowax ➢ REMEDY: → Add (?) in waterbath → Use floating solutions: (?)

A

soap or 10% polyethylene glycol 900

Pearse solution or Blank and McCarthy solution

57
Q

✓ Purified pyroxylin nitrocellulose

A

CELLOIDIN PROCESSING

58
Q

✓ Suitable for specimens containing large cavities or hollow spaces which tend to collapse (eyes) & for larger embryos

A

CELLOIDIN PROCESSING

59
Q

✓ Available in thin (2%), medium (4%) and thick (8%) solutions

A

CELLOIDIN PROCESSING

60
Q

✓ DIS: Tissues cannot be cut as thin as they are with paraffin wax

A

CELLOIDIN PROCESSING

61
Q

✓ ADV: Causes much less shrinkage & distortion; slow process

A

CELLOIDIN PROCESSING

62
Q

➢ With rubbery consistency – w/o distortion

A

CELLOIDIN PROCESSING

63
Q

➢ Does not require heat

A

CELLOIDIN PROCESSING

64
Q

➢ Soluble in equal concentrations of ether and alcohol

A

Low Viscosity Nitrocellulose

65
Q

✓ Can be used in higher concentrations

A

Low Viscosity Nitrocellulose

66
Q

✓ Produces harder tissue blocks

A

Low Viscosity Nitrocellulose

67
Q

o Cracking of tissue sections and chrome-mordanted tissues may crumble

A

Low Viscosity Nitrocellulose

68
Q

→ REMEDY: add plasticizers like Castor oil, Oleum ricini

A

Low Viscosity Nitrocellulose m

69
Q

o More explosive than celloidin

A

Low Viscosity Nitrocellulose

70
Q

o Cannot be stored

A

Low Viscosity Nitrocellulose

71
Q

❖ for bones, teeth large brain sections & whole organs

A

WET METHOD

72
Q

WET METHOD ❖ Fixation → Dehydration → Equal parts ether and alcohol (12-24 hours) → Thin celloidin: 2-4 % (5-7 days) → Medium celloidin: 4-6 % (5-7 days) → Thick celloidin: 812 % (3-5 days) → Embedding: fresh thick celloidin in a jar or desiccator (fingerprint no longer leaves a mark on block surface) → Storage: 70-80% alcohol

A
73
Q

❖ Same with wet method EXCEPT for the following steps

A

DRY METHOD

74
Q

✓ Gilson’s mixture (chloroform + cedarwood oil) is added to the fresh thick celloidin during embedding

A

DRY METHOD

75
Q

✓ Storage in alcohol is contraindicated

A

DRY METHOD

76
Q

❖ Provided superior results for light microscopic studies especially of hard tissues and samples for high resolution microscopy

A

PLASTICS

77
Q
  • slowest
A

Araldite base (bisphenol)

78
Q
  • fastest
A

Cyclohexene Dioxide (spurr)

79
Q

➢ Reduce antigenicity

A

EPOXY

80
Q

➢ Sensitization on skin contact and inhalation

A

EPOXY

81
Q

➢ Contains toxic components (eg. Vinylcyclohexane dioxide)

A

EPOXY

82
Q

➢ For high resolution LM

A

ACRYLIC

83
Q

: valued for its hydrophilic nature

A

➢ Glycol Methacrylate (GMA)

84
Q

: valued for its hardness

A

➢ Methyl Methacrylate (MMA)

85
Q

– added as catalyst

A

➢ Benzoyl Peroxide

86
Q

→ acts as an active site (polymerization of acrylics)

A

➢ Benzoyl Peroxide

87
Q

✓ Histochemical and enzyme studies

A

GELATIN PROCESSING

88
Q

✓ Delicate specimens

A

GELATIN PROCESSING

89
Q

✓ Frozen sections

A

GELATIN PROCESSING

90
Q

✓ Water-soluble

A

GELATIN PROCESSING

91
Q

✓ Low MP and does not overharden

A

GELATIN PROCESSING

92
Q

GELATIN PROCESSING ❖ Fixation → washing → 10 % (24 hours) → 20 % (12 hours) → 20% until impregnation and embedding are complete → 10 % formalin (12-24 hours)

A
93
Q

**All gelatin reagents contain

A

1 % phenol

94
Q

✓ Arranging the tissue in the mold:

A

embedding

95
Q

✓ Fixing the tissue block on the microtome:

A

prior to sectioning

96
Q

✓ Fixing the tissue block on the microtome:

A

prior to sectioning

97
Q

✓ Arranging the tissue ribbons on the slide:

A

flotation

98
Q

✓ 2 L-shaped heavy metal arranged flat on a metal surface

A
  1. Leuckhart’s embedding mold
99
Q

✓ with several interlocking plates making several compartments

A
  1. Compound E unit
100
Q

✓ ADV: cheap, diff sizes, avoids confusion

A

Paper boats

MTAP - HISTOPATHOLOGIC & CYTOLOGIC TECHNIQUES MODULE (9 decks)

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